Allogeneic cell therapy products, such as therapeutic cells derived from pluripotent stem cells (PSCs), have amazing potential to treat a wide variety of diseases and vast numbers of patients globally. However, there are various challenges related to manufacturing PSCs in single-use bioreactors, particularly at larger volumetric scales. This manuscript addresses these challenges and presents potential solutions to alleviate the anticipated bottlenecks for commercial-scale manufacturing of high-quality therapeutic cells derived from PSCs.

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Cell Culture Process Scale-Up Challenges for Commercial-Scale Manufacturing of Allogeneic Pluripotent Stem Cell Products

The demand for large cell numbers for cellular therapies and drug screening applications requires the development of scalable platforms capable of generating high-quality populations of tissue-specific cells derived from human pluripotent stem cells (hPSCs). Here, we studied the ability of Gibco StemScale PSC Suspension Medium to promote the efficient expansion of hPSC cultures as aggregates grown in suspension. We tested human induced pluripotent stem cell (hiPSC) growth in 6-well plates (on orbital shaker platforms) and single-use vertical-wheel bioreactors for a total of three consecutive passages. Up to a 9-fold increase in cell number was observed over 5 days per passage, with a cumulative fold change up to 600 in 15 days. Additionally, we compared neural induction of hiPSCs by using a dual SMAD inhibition protocol with a commercially available neural induction medium, which can potentially yield more than a 30-fold change, including neural progenitor induction and expansion. This system can also be adapted toward the generation of floor plate progenitors, which yields up to an 80-fold change in cell number and generates FOXA2-positive populations. In summary, we developed platforms for hiPSC expansion and neural induction into different brain regions that provide scalability toward producing clinically relevant cell numbers.

A Dynamic 3D Aggregate-Based System for the Successful Expansion and Neural Induction of Human Pluripotent Stem Cells

Correspondence Sunghoon Jung, PBS Biotech Inc., 4721 Calle Carga, Camarillo, CA, 93012 USA. Email: sjung@pbsbiotech.com Abstract The purpose of this investigation was to develop an initial set of Power number versus Reynolds number results for a family of vertical-wheel bioreactors. These bioreactors are increasingly being used for the manufacture of cells for cell therapy but have not been characterized according to this approach. A novel gravimetric method to measure power was used, and the validity of this method was assessed by measuring power for a standard stirred tank bioreactor with a Rushton impeller. The results of the gravimetric method were found to closely match those derived from traditional methods. The validated method was then used to measure the power draw and develop an initial set of Power number versus Reynolds number results for a family of vertical-wheel bioreactors.

Initial Power Measurements for a Family of Novel
 Vertical‐Wheel
 Bioreactors

Preface to the special issue honouring Professor Leo A. Behie

Generation of pure pancreatic progenitor cells (PPs) is critical for clinical translation of stem cell derived islets. Herein, we performed PP differentiation with and without AKT/P70 inhibitor AT7867 and characterized the resulting cells at protein and transcript level in vitro and in vivo upon transplantation into diabetic mice. AT7867 treatment increased the percentage of PDX1+NKX6.1+ (-AT7867: 50.9% [IQR 48.9%-53.8%]; +AT7867: 90.8% [IQR 88.9%-93.7%]; p=0.0021) and PDX1+GP2+ PP cells (-AT7867: 39.22% [IQR 36.7%-44.1%; +AT7867: 90.0% [IQR 88.2%-93.6%]; p=0.0021). Transcriptionally, AT7867 treatment significantly upregulated PDX1 (p=0.0001), NKX6.1 (p=0.0005) and GP2 (p=0.002) expression compared to controls, while off-target markers PODXL (p<0.0001) and TBX2 (p <0.0001) were significantly downregulated. Transplantation of AT7867 treated PPs resulted in faster hyperglycemia reversal in diabetic mice compared to controls (time and group: p<0.0001). Overall, our data shows that AT7867 enhances PP cell differentiation leading to accelerated diabetes reversal.

/pdf/at7867-promotes-pancreatic-progenitor-differentiation-of-3bir9cn8.pdf

AT7867 promotes pancreatic progenitor differentiation of human iPSCs and accelerates diabetes reversal

Computational fluid dynamic characterization of vertical-wheel bioreactors used for effective scale-up of human induced pluripotent stem cell aggregate culture

Shivek Kanwar

Papers

Computational fluid dynamic characterization of vertical-wheel bioreactors used for effective scale-up of human induced pluripotent stem cell aggregate culture